Programa

SEMESTER 1

1. Pluripotency control mechanisms during development

An important feature of organs, is their ability to maintain their morphology and function in spite of environmental or genetic stimuli that cause natural or accidental cell loss. This capacity is often sustained by the so-called stem cells, which are able to provide new cells of different types in each tissue.

A common characteristic shared by these pluripotent cells, is their ability to remain in a “dormant” status while being able at any given point to activate their regenerative potential. The mechanisms controlling this ability have to be precisely regulated while balancing between the need to maintain tissues’ homeostasis and having to retain the limit of uncontrolled cellular growth. In mechanistic terms, this delicate balance is sustained during development by a continuous feedback loop between metabolic shifts, response to hormonal cues and regulation of gene expression.

Drosophila melanogaster, is a model organism used extensively over the past decades in several developmental studies. The well structured knowledge we now have in several aspects of the Drosophila’s biology, allows us to use this organism as an in vivo model in several studies, including the functional characterization and the dissection of the molecular mechanisms that govern the regulation of stem cells.

In this course, we will take advantage of the powerful Drosophila genetic tools to mark progenitor cells and modify the expression levels of selected genes that regulate some of their distinct characteristics. This will give us the opportunity to study basic mechanisms that keep these cells in a multi-potent status and identify crucial aspects of their role throughout the development.

2. The code of life

The information encoded in the genetic material is translated into proteins following a set of rules known as the Genetic Code, which is universal for almost all the living organisms and establishes the basis for life. This means that, from the simplest bacterium to us, we all use a highly similar “language” to synthesize all the required proteins, a language based on the combination of four nucleotides (A, U, G and C). The combination of these nucleotides in groups of three (the codons) represent the “words” that all living organisms “read” in the ribosomes to elongate a chain of amino acids, eventually generating proteins.

Malfunctions of the different biomolecules participating in this highly regulated system can lead to disease, but in recent years strategies to modify some of their functions have been applied for biotechnological purposes.

In this course we will study the role of the different key constituents of the translation machinery in cells, and the essential importance of the Genetic Code in the generation of active proteins. At the practical side, we will learn the common laboratory techniques used for protein expression and purification in a prokaryotic system (Escherichia coli), and also certain specific techniques to introduce mutations in the sequence of genes to produce recombinant proteins with modified functions.

3. Chemical synthesis of bioactive molecules

Chemistry is at the heart of the drug-discovery process. For each new drug reaching the market, there have been billions of compounds synthesised and tested. Why so many? Because is by chemically modulating the structure of the molecules that we can fine-tune their potency and also their pharmacodynamic/pharmacokinetic properties (absorption, distribution, metabolism, excretion and side-effects).

In this practical course we will synthesise acetaminophen, also known as paracetamol or the active ingredient in Gelocatil®. While doing so we will learn how chemists plan and design the synthesis routes as well as the main techniques and instruments used to build the active ingredients of all small-molecule drugs.

4. Meet the guardians of the DNA

Our DNA can break due to external (i.e. the sun and chemical agents) and internal (i.e. DNA replication) factors. To solve this problem, we have some genes that function as the guardians of our genome, by identifying these lesions and activating the DNA Damage Response (DDR) pathway. However, the DDR pathway does not always work properly. If the damage is severe, the cell undergoes to apoptosis (programmed cell death) or senescence (permanent cell cycle arrest). On the other hand, if the stability of the genome is not massively compromised, the cell survives and starts proliferating in an uncontrolled manner. The transformation of a normal cell into a tumour cell is a multi-step process that involves dynamic changes in the genome at both the genetic and epigenetic levels. Currently, cancer accounts for 8 million deaths per year, being the most common cause of death in Europe after cardiovascular diseases.

Our lab is interested in deciphering the potential role of genomic instability and the DDR in cancer predisposition. In this practical course, we will monitor the effect of DNA damage in different cancer cell lines and perform some techniques that are daily used in the lab to study this phenomenon.

Colorectal Cancer (CRC) is the most common cancer and the second leading cause of cancer-related deaths in Spain, according to the AECC (Asociación Española Contra el Cáncer). CRC produces cancers in the large intestine through a multi-step process that, like many other types of cancer, starts with the acquisition of mutations in key genes. These mutations then go on to make cells proliferate without control. CRC can be easily treated if detected early. However, we still need to find effective treatments for the most advanced stages of cancer, particularly for metastasis—the final and deadliest step of cancer, which in the case of CRC occurs mainly in the liver.

The great potential of cancer cells resides not only in their capacity for mass proliferation but also in their ability to trick the healthy cells surrounding them. Fibroblasts, cells of the immune system, and blood vessels form what we call the tumour microenvironment (TME), which protects cancer cells from being removed and facilitates their invasion of other healthy organs. Our laboratory has recently shown that the TME is crucial for the aggressiveness of CRC and formation of metastasis in the liver. We are now channelling efforts into studying the potential of the TME as a way to block metastasis and into identifying novel treatments.

In this practical course we will provide an overview of several in vivo models and histological tools. Using these tools, we will discover and target those cells that form the TME of liver metastasis, with the aim to learn how to exploit them and stop the CRC metastasis.

6. Targeting cell signaling pathways to treat cancer

Cells have to constantly deal with changes in their extracellular environment. It means that they have to respond efficiently to these changes in order to prevent damage but also they have to control proliferation, survival and migration. To do that, cells have developed several mechanisms that allow them to receive, integrate and interpret signals to produce the appropriate response. These mechanisms, called ‘signaling pathways’, are based in a series of chemical changes. One of the most important modifications is phosphorylation and proteins which are able to phosphorylate others are called kinases.

Our laboratory is mainly focused in a particular protein kinase called p38 MAPK, which is activated under different stress situations and plays a critical role in inflammation, cell growth, proliferation, differentiation and cell death. However, when this pathway is dysregulated diseases such as cancer can arise. In that case, p38 MAPK inhibition could be beneficial to treat patients.

In this practical course we will provide an overview of the p38 MAPK signaling pathway and which are the main techniques we use in the lab to study it. We will use purified proteins and cultured cell lines to learn how p38 MAPK inhibitors work. Finally, we will explore which patients could benefit from them.

SEMESTER 2

1. The Role of Microtubules in Cell Division and Cancer

Cellular cytoskeleton is perhaps one of the most important structures of the cell as it is involved in several functions as morphogenesis, cellular division or cell migration. It is composed by three members, actin filaments, intermediate filaments and microtubules. Microtubules are hollow cylindric polymers composed by tubulin dimers. Microtubules are very dynamic structures that constantly undergo episodes of polymerization and depolimerization which is known as dynamic instability. Microtubules are involved in key processes as intracellular transport of cargos (proteins, vesicles and even organelles) and in mitosis where they are in charge of the formation of the mitotic spindle and proper chromosome segregation. One of the hallmarks of cancer is uncontrolled cell division and one approach to treat cancer is to try to kill cancer cells. Compounds that target microtubules disrupting the normal cell division cycle and eventually lead to cell death have been proven to be one of the most effective cancer chemotherapeutic drugs available.

In this practical course, we will learn how a mitosis occurs in vivo, what are the different phases of mitosis, what is the role of microtubules in this process and how can we treat cancer focusing on microtubules. For this purpose, we will use cultured cell lines treated with several chemotherapeutic drugs used in cancer therapy and see how they affect cell division with the help of fluorescence light microscopy techniques.

In the year 2000, the Human Genome Project was completed at a cost of around 3 billion dollars and ten years of intense work. The main conclusion was that over 98% of our genetic material was junk DNA. Far from resolving our questions, by reading our genome it became apparent that we didn't understand it. Since then, mounting evidence has made us realise that understanding our genome means understanding how its expression is regulated.

We are now starting to gain some insight into how cells depend on extremely complex regulatory networks to switch genes on and off through a myriad of novel players such as microRNAs, long non-coding RNAs, and RNA-binding proteins. Unravelling the mechanisms underlying the regulation of gene expression will be vital to understand how an erythrocyte and a neuron differ while sharing the exact same genes or why in identical twins one falls ill with a disease like cancer while the other remains completely healthy.

In this course we will use in vitro cell culture systems and look at the current methods that researchers apply to study the regulation of gene expression.

3. Strategies to understand ageing and senescence

Ageing is characterised by a gradual deterioration of physiological function, and it is considered the primary risk for human pathologies such as cancer or cardiovascular disorders. One of the mail hallmarks of ageing is cellular senescence, the phenomenon by which normal cells cease to divide. Senescent cells accumulate in the organism over time, secreting pro-inflammatory molecules and contributing to ageing-related diseases.

There is an increasing interest in clinical medicine to better identify and target senescent cells, as their elimination delays and ameliorates some ageing-associated diseases, and can even extend longevity.

Students will acquire hands-on experience using different techniques to induce cellular senescence in normal cells. They will learn state of the art techniques to specifically target senescent cells, analysing in vitro and in vivo samples. Finally, they will also have the opportunity to perform classical protocols to detect senescent cells such as the SAβgal staining. These techniques will help us to understand, identify and target senescent cells, which is a promising therapeutic approach in clinical medicine.

The most common type of human tumor is the carcinoma. Carcinoma is derived from epithelial tissue, such as the skin, but it can become invasive or metastatic by spreading beyond the primary tissue layer to surrounding tissues or organs. These transition is mediated by the activation, in aggressive cancer cells, of the EMT programme (Epithelial to Mesenchymal Transition) causing them to undergo morphogenetic alterations that allows a higher invasion capacity.

The fruit fly, Drosophila melanogaster, with a long and impressive history as a model organism for studying epithelial development, revealed itself as a potent tool for studying carcinomas offering the possibility of analyzing the detailed interaction between cells, tissues and genes. Due to the fact that this fly maintains conserved the signaling pathways present in humans, we also have the possibility of reducing genome redundancy when establishing comparations between the two.

In this practical course, we will learn about Drosophila has a model organism for studying cancer related mechanisms. We will identify different fly structures such as the wing imaginal disc and the fat body through in vivo dissections and perform immunohistochemistry in order to understand how different signaling pathways are analyzed.

The cellular membrane separates the interior of the cell from the outside environment. To allow a selective permeability and another cellular functions, this membrane contents numerous proteins. These molecules are integrated in the membrane and their functions include signaling, enzymatic activity, cell–cell contact, cytoskeleton contact, surface recognition, or transporting substances across the membrane. Due to the numerous functions, mutations or abnormalities in their expression levels cause different pathologies or physiological dysfunctions such as cancer, intolerances, hormonal imbalances or certain rare diseases.

To facilitate the functional and structural study of membrane proteins, the molecule of interest is overexpressed heterologously in an expression system, the cellular membrane is purified, next the protein is extracted from the membrane by using detergents, and lately it is purified.

In this practical course we will learn how the cellular membrane is isolated from yeast (Pichia pastoris) and mammal cells expressing a human amino acid transporter involved in several pathologies. Among the most used expression systems for membrane proteins, mammal and yeast cells are found. In addition, we will see the advantages and disadvantages of both used expression systems in regard to the recombinant protein production for functional or structural studies.

Cancers is a disease where cells start to grow and proliferate in an uncontrolled way. These cells accumulate somewhere from a few to millions of changes in their DNA, which are called somatic mutations. But only a subset of them confer malignant properties (driver mutations) whereas most have little or no impact on how the cell functions (passenger mutations). The finding of driver mutations is essential for understanding the biology of cancer and developing the so-called precision medicine, which relies on identifying the driver mutations of the patient to prescribe drugs specifically targeting them. We call this process in silico drug prescription.

Analyzing thousands of tumors with all their mutations is a challenging task –luckily, we can use computers to help us analyze and interpret this vast amount of biological data (bioinformatics!). During the program the students will be introduced to the fundamentals of Bioinformatics and Cancer Genomics. They will use and write several programs in the Python programming language to analyze the mutations of thousands of tumors from real patients, and perform a state-of-the-art in silico drug prescription according to the driver mutations found. They will also be introduced to the most common forms of visualization of cancer mutational data. No prior programming skills are required.